Thiamine phosphate | 532-40-1

• 分子结构分类: 有机化合物 - 有机杂环化合物 - 二嗪类
• 英文名称: Thiamine phosphate
• 英文别名: 2-[3-[(4-amino-2-methylpyrimidin-5-yl)methyl]-4-methyl-1,3-thiazol-3-ium-5-yl]ethyl phosphate
• CAS: 532-40-1
• 化学式: C₁₂H₁₆N₄O₄PS
• 分子量: 343.323
• InChiKey: HZSAJDVWZRBGIF-UHFFFAOYSA-M

物化性质

• 熔点：
  ~200°
• 密度：
  1.53[at 20℃]
• 溶解度：
  甲醇（微溶）、水（微溶）
• LogP：
  -4.7 at 20℃

计算性质

• 辛醇/水分配系数(LogP)：
  -0.2
• 重原子数：
  22
• 可旋转键数：
  5
• 环数：
  2.0
• sp3杂化的碳原子比例：
  0.42
• 拓扑面积：
  156
• 氢给体数：
  1
• 氢受体数：
  8

反应信息

• 作为反应物：
  描述：

  水 、 Thiamine phosphate 生成 H3PO4 、 硫胺素
  参考文献：
  名称：

  A th-1 Mutant of Arabidopsis thaliana Is Defective for a Thiamin-Phosphate-Synthesizing Enzyme: Thiamin Phosphate Pyrophosphorylase

  摘要：

  我们研究了阿拉伯芥野生型和突变型（th-1）中硫胺素磷酸酯激酶的活性，突变型需要外源硫胺素才能生长。在1×10-7摩尔硫胺素中生长的突变型和野生型植物被用于研究硫胺素和硫胺素单磷酸（TMP）的产生，使用4-甲基-5-羟乙基噻唑磷酸盐和2-甲基-4-氨基-5-羟甲基嘧啶焦磷酸盐作为底物。野生型菌株同时形成硫胺素和TMP，而th-1突变型则不能。当TMP被添加到提取物中时，th-1突变型和野生型都会产生硫胺素。因此，可以得出结论，th-1突变型在TMP焦磷酸酶的活性方面存在缺陷。对野生型植物中酶的一些特性进行了研究。反应的最佳温度是45°C，底物的Km值是4-甲基-5-羟乙基噻唑磷酸盐的2.7×10-6摩尔和2-甲基-4-氨基-5-羟甲基嘧啶焦磷酸盐的1.8×10-6摩尔。

  DOI：

  10.1104/pp.88.2.248
• 作为产物：
  描述：

  4-Methyl-5-(2-phosphonatooxyethyl)thiazole(2-) 、 4-Amino-2-methyl-5-(diphosphooxymethyl)pyrimidine 生成 Thiamine phosphate
  参考文献：
  名称：

  Biosynthesis of Thiamin I: The Function of the thiE Gene Product

  摘要：

  DOI：

  10.1021/ja00113a025

文献信息

• A <i>th-1</i> Mutant of <i>Arabidopsis thaliana</i> Is Defective for a Thiamin-Phosphate-Synthesizing Enzyme: Thiamin Phosphate Pyrophosphorylase
  作者：Yoshibumi Komeda、Miyako Tanaka、Takahiro Nishimune

  DOI：10.1104/pp.88.2.248

  日期：1988.10.1

  We have examined the activity of the thiamin phosphate pyrophosphorylase in Arabidopsis thaliana wild type and in a mutant (th-1) which requires exogenous thiamin for growth. Mutant and wild-type plants grown in 1 × 10−7 molar thiamin were used for the examination of the production of thiamin and thiamin monophosphate (TMP) using 4-methyl-5-hydroxyethylthiazole phosphate and 2-methyl-4-amino-5-hydroxymethylpyrimidine pyrophosphate as substrates. While the wild-type strain formed both thiamin and TMP, the th-1 mutant did not. When TMP was added to the extracts, the th-1 mutant, as well as wild type, produced thiamin. Accordingly, it was concluded that the th-1 mutant was defective in the activity of TMP pyrophosphorylase. Some of the characteristics of the enzyme from the wild-type plant were examined. The optimum temperature for the reaction is 45°C, and the Km values for the substrates are 2.7 × 10−6 molar for 4-methyl-5-hydroxyethylthiazole phosphate and 1.8 × 10−6 molar for 2-methyl-4-amino-5-hydroxymethylpyrimidine pyrophosphate.

  我们研究了阿拉伯芥野生型和突变型（th-1）中硫胺素磷酸酯激酶的活性，突变型需要外源硫胺素才能生长。在1×10-7摩尔硫胺素中生长的突变型和野生型植物被用于研究硫胺素和硫胺素单磷酸（TMP）的产生，使用4-甲基-5-羟乙基噻唑磷酸盐和2-甲基-4-氨基-5-羟甲基嘧啶焦磷酸盐作为底物。野生型菌株同时形成硫胺素和TMP，而th-1突变型则不能。当TMP被添加到提取物中时，th-1突变型和野生型都会产生硫胺素。因此，可以得出结论，th-1突变型在TMP焦磷酸酶的活性方面存在缺陷。对野生型植物中酶的一些特性进行了研究。反应的最佳温度是45°C，底物的Km值是4-甲基-5-羟乙基噻唑磷酸盐的2.7×10-6摩尔和2-甲基-4-氨基-5-羟甲基嘧啶焦磷酸盐的1.8×10-6摩尔。
• Systematic discovery of analogous enzymes in thiamin biosynthesis
  作者：Enrique Morett、Jan O Korbel、Emmanuvel Rajan、Gloria Saab-Rincon、Leticia Olvera、Maricela Olvera、Steffen Schmidt、Berend Snel、Peer Bork

  DOI：10.1038/nbt834

  日期：2003.7

  occurrences of genes across species. Our approach, applied to the thiamin biosynthesis pathway comprising approximately 15 catalytic steps, predicts seven instances in which known enzymes have been displaced by analogous proteins. So far we have verified four predictions by genetic complementation, including three proteins for which there was no previous experimental evidence of a role in the thiamin biosynthesis

  迄今为止，在所有已完成的基因组测序项目中，各个物种的生化途径中均发现了相当数量的“缺口”。在许多情况下，缺失的酶被类似物，功能等效的蛋白质取代，这些蛋白质已经独立进化且缺乏序列和结构相似性。在这里，我们通过分析物种间基因的反相关出现来填补这些空白。我们的方法应用于包含约15个催化步骤的硫胺素生物合成途径，预测了7种实例中已知酶已被类似蛋白取代的情况。到目前为止，我们已经通过遗传互补验证了四个预测，包括三个蛋白，以前没有实验证据证明这些蛋白在硫胺素生物合成途径中起作用。对于一种假设的蛋白质，生化特征证实了预测的硫胺素磷酸合酶（ThiE）活性。结果证明了我们的计算方法能够预测特定功能，而无需考虑序列相似性。
• Identification of Inhibitors against Mycobacterium tuberculosis Thiamin Phosphate Synthase, an Important Target for the Development of Anti-TB Drugs
  作者：Garima Khare、Ritika Kar、Anil K. Tyagi

  DOI：10.1371/journal.pone.0022441

  日期：——

  Tuberculosis (TB) continues to pose a serious challenge to human health afflicting a large number of people throughout the world. In spite of the availability of drugs for the treatment of TB, the non-compliance to 6–9 months long chemotherapeutic regimens often results in the emergence of multidrug resistant strains of Mycobacterium tuberculosis adding to the precariousness of the situation. This has necessitated the development of more effective drugs. Thiamin biosynthesis, an important metabolic pathway of M.tuberculosis, is shown to be essential for the intracellular growth of this pathogen and hence, it is believed that inhibition of this pathway would severely affect the growth of M.tuberculosis. In this study, a comparative homology model of M.tuberculosis thiamin phosphate synthase (MtTPS) was generated and employed for virtual screening of NCI diversity set II to select potential inhibitors. The best 39 compounds based on the docking results were evaluated for their potential to inhibit the MtTPS activity. Seven compounds inhibited MtTPS activity with IC50 values ranging from 20 – 100 µg/ml and two of these exhibited weak inhibition of M.tuberculosis growth with MIC99 values being 125 µg/ml and 162.5 µg/ml while one compound was identified as a very potent inhibitor of M.tuberculosis growth with an MIC99 value of 6 µg/ml. This study establishes MtTPS as a novel drug target against M.tuberculosis leading to the identification of new lead molecules for the development of antitubercular drugs. Further optimization of these lead compounds could result in more potent therapeutic molecules against Tuberculosis.

  结核病（TB）仍然对人类健康构成严重威胁，影响着全世界大量人口。尽管有治疗结核病的药物，但患者往往不遵守长达6-9个月的化疗方案，导致耐多药结核杆菌的出现，使情况更加危险。因此有必要开发更有效的药物。硫胺素生物合成是结核杆菌的重要代谢途径，对这种病原体的细胞内生长至关重要，因此抑制该途径将严重影响结核杆菌的生长。在这项研究中，我们构建了结核杆菌硫胺素磷酸合成酶（MtTPS）的同源性比较模型，并用于NCI多样性集II的虚拟筛选，以选择潜在的抑制剂。根据对接结果，我们对39种最佳化合物进行了评估，以确定其抑制MtTPS活性的潜力。其中7种化合物抑制MtTPS活性的IC50值在20-100微克/毫升之间，其中两种化合物对结核杆菌的生长表现出弱抑制作用，MIC99值分别为125微克/毫升和162.5微克/毫升，而一种化合物被确定为结核杆菌生长的强效抑制剂，MIC99值为6微克/毫升。这项研究将MtTPS确定为抗结核杆菌的新药靶点，为抗结核药物的开发提供了新的先导分子。对这些先导化合物的进一步优化可能会产生更有效的抗结核治疗分子。
• Copurification of hydroxyethylthiazole kinase and thiamine-phosphate pyrophosphorylase of Saccharomyces cerevisiae: characterization of hydroxyethylthiazole kinase as a bifunctional enzyme in the thiamine biosynthetic pathway
  作者：Y Kawasaki

  DOI：10.1128/jb.175.16.5153-5158.1993

  日期：1993.8

  Mutants of Saccharomyces cerevisiae resistant to 2-amino-4-methyl-5-beta-hydroxyethylthiazole, an antimetabolite of 4-methyl-5-beta-hydroxyethylthiazole (hydroxyethylthiazole), which are deficient in the activities of both hydroxyethylthiazole kinase and thiamine-phosphate pyrophosphorylase, involved in the pathway of de novo synthesis of thiamine in S. cerevisiae, have been isolated. Genetic analysis revealed that the mutation occurs at a single gene in the nucleus. The two enzyme activities were copurified to apparent homogeneity, and the molecular masses of the purified proteins were found to be approximately 470 and 60 kDa, as determined by gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, respectively. Hydroxyethylthiazole kinase was specific for ATP and Mg2+, although to a lesser extent a combination with other nucleoside triphosphates or divalent cations could replace them. p-Chloromercuribenzoate was a potent inhibitor of the enzyme, and the inhibition was prevented by the addition of 2-mercaptoethanol. These findings indicate that yeast hydroxyethylthiazole kinase is a bifunctional enzyme with thiamine-phosphate pyrophosphorylase activity, which is an octamer of identical 60-kDa subunits.

  孟德酵母突变体对2-氨基-4-甲基-5-β-羟乙基噻唑酮（4-甲基-5-β-羟乙基噻唑酮（羟乙基噻唑酮）的一种抗代谢物）具有抗性，这些突变体缺乏羟乙基噻唑酮激酶和硫胺酸磷酸二酯酶的活性，这些酶涉及S. cerevisiae中硫胺素新生途径。遗传分析表明，该突变发生在细胞核的单个基因中。两种酶活性被共纯化到表观同质性，并且纯化蛋白的分子量分别为约470和60 kDa，由凝胶过滤和SDS-PAGE测定。羟乙基噻唑酮激酶对ATP和Mg2 +具有特异性，虽然其他核苷酸三磷酸盐或二价阳离子的组合可以代替它们，但程度较小。对羟乙基噻唑酮激酶的强效抑制剂是对氯汞苯甲酸，而加入2-巯基乙醇可以防止该抑制。这些发现表明，酵母中的羟乙基噻唑酮激酶是一种具有硫胺酸磷酸二酯酶活性的双功能酶，是相同的60 kDa亚基的八聚体。
• Characterization of the Bacillus subtilis thiC operon involved in thiamine biosynthesis
  作者：Y Zhang、S V Taylor、H J Chiu、T P Begley

  DOI：10.1128/jb.179.9.3030-3035.1997

  日期：1997.5

  The characterization of a three-gene operon (the thiC operon) at 331 min, which is involved in thiamine biosynthesis in Bacillus subtilis, is described. The first gene in the operon is homologous to transcription activators in the lysR family. The second and third genes (thiK and thiC) have been subcloned and overexpressed in Escherichia coli. ThiK (30 kDa) catalyzes the phosphorylation of 4-methyl-5-(beta-hydroxyethyl)thiazole. ThiC (27 kDa) catalyzes the substitution of the pyrophosphate of 2-methyl-4-amino-5-hydroxymethylpyrimidine pyrophosphate by 4-methyl-5-(beta-hydroxyethyl)thiazole phosphate to yield thiamine phosphate. Transcription of the thiC operon is not regulated by thiamine or 2-methyl-4-amino-5-hydroxymethylpyrimidine and is only slightly repressed by 4-methyl-5-(beta-hydroxyethyl)thiazole.

  本文描述了位于331分钟处的三基因操纵子（thiC操纵子），该操纵子参与枯草杆菌中硫胺素生物合成的特征。操纵子中的第一个基因与溶菌酶家族的转录激活因子同源。第二个和第三个基因（thiK和thiC）已经在大肠杆菌中进行了亚克隆和过度表达。 ThiK（30 kDa）催化4-甲基-5-（β-羟乙基）噻唑的磷酸化。ThiC（27 kDa）催化2-甲基-4-氨基-5-羟甲基嘧啶焦磷酸二酯的焦磷酸基被4-甲基-5-（β-羟乙基）噻唑磷酸酯取代，生成硫胺素磷酸酯。 thiC操纵子的转录不受硫胺素或2-甲基-4-氨基-5-羟甲基嘧啶的调节，并且仅受到4-甲基-5-（β-羟乙基）噻唑的轻微抑制。